Extended Data Fig. 2: Analysis of ISC components by FIDA.
From: Cross-regulation of [2Fe–2S] cluster synthesis by ferredoxin-2 and frataxin
a, Illustration of the experimental principle of FIDA. The capillary is first equilibrated with an analyte, followed by injection of the fluorescently labelled indicator (adapted from ref. 54 under a Creative Commons licence CC BY 3.0). Indicator flow along the capillary is induced and the fluorescent profile is measured at the detector window. b, Kinetics of [2Fe–2S] cluster synthesis by the recombinant ISC system using FXN and FDX2 either unlabelled (FXN, FDX2) or labelled (FXNALC, FDX2ALC). c, FIDA Taylorgram profiles of FXNALC and FDX2ALC alone or after saturation with the (NIAU)2 complex. d,e, Corresponding hydrodynamic radius of labelled proteins and their complexes determined by FIDA mapped as a sphere alongside the structure of FXN (PDB: 6NZU) and the FXN–(NIAU)2 complex (PDB: 6NZU) (d) or FDX2 (AlphaFold v.2.2) and the FDX2–(NIAU)2 complex (AlphaFold v.2.2) (e). f, FIDA competition assays of unlabelled oxidized (blue) and reduced (red) FDX2 titrated into a preformed FXNALC–(NIAU)2 complex. (b, representative data selected from n = 3 independent experiments with similar results; f, mean, ± s.d., n = 3 independent experiments).
